计算溶液所需的质量、体积或浓度。
,适用于对基线干扰要求严格的色谱和分析工作流程。
-20°C储存。超低温运输 。请查阅批次 COA 获取详细规格。
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在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
本制品是经过大肠杆菌表达的重组蛋白,与RNase A形成 1:1 复合体,对RNase表现高度的非竞争性抑制,本品属蛋白质性质,与其它竞争性抑制剂(核酸类、无机磷酸类)不同,可以很容易地通过苯酚处理将其从反应体系中除去。
产品内容
RNase Inhibitor(RNasin)(40U/μl)
纯度
1)300U的RNasin和1μg的λDNA-Hind III分解物在37℃下反应1小时,DNA的电泳谱带不发生变化。
2)300U的RNasin和 1μg的Closed circμlar(RFI)pBR322 DNA 在37℃下反应1小时,DNA的电泳谱带不发生变化。
3)100U的RNasin和 1μg的16S,23S rRNA在37℃下反应1小时,RNA的电泳谱带不发生变化。
4)SDS-PAGE:在分子量64KDa处是单一带。
应用
1)cDNA 合成反应;
2)多核糖体的分离(Polysome isolation);
3)体外翻译(In vitro translation);
4)SP6 或 T7 RNA 聚合酶的体外转录
5)体外无细胞系统转录(In vitro transcription with cell-free extract)。
使用注意
1)抑制活性的 pH 值范围较广,在 pH7~8 时表现最大活性;
2)起泡或强烈搅拌(Vortex 等)会引起失活;
3)不抑制 RNase H 活性。
使用例
cDNA 第一条链的合成。
在微量离心管中配制下列反应液,全量为20μl
试剂 | 使用量 | 终浓度 |
Total RNA or Poly(A)RNA | 1-5μg | — |
5×MMLV RT Buffer | 4μl | 1× |
dNTP Mixture(25mM ) | 0.4μl | 0.5mM |
Oligo dT or Random Primer (20μM) | 1μl | 1μM |
RNasin(40U/μl) | 1μl | 2U/μl |
MMLV Reverse Transcriptase(200U/μl) | 1μl | — |
超纯水 | Up to 20μl | — |
总体积 | 20μl | — |
1) 轻搅拌。
3) 室温下10min放置后,移入42℃温槽中。
4) 42℃保温1小时。
4) 把样品移入冰中冷却2min。
This product is a recombinant protein expressed in Escherichia coli and forms a 1:1 complex with RNase A. It exhibits a high degree of non-competitive inhibition of RNase. This product is of protein nature and is compatible with other competitive inhibitors (nucleic acids, inorganic phosphates). ) Different, it can be easily removed from the reaction system by phenol treatment.
Product content
RNase Inhibitor (RNasin) (40U/μl)
Purity
1) 300 U of RNasin and 1 μg of λDNA-Hind III decomposition product were reacted at 37°C for 1 hour, and the DNA electrophoresis band did not change.
2) 300U RNasin and 1μg Closed circμlar (RFI) pBR322 DNA are reacted at 37°C for 1 hour, and the DNA electrophoresis band does not change.
3) 100 U of RNasin and 1 μg of 16S, 23S rRNA are reacted at 37°C for 1 hour, and the electrophoresis band of RNA does not change.
5) SDS-PAGE: a single band at a molecular weight of 64KDa.
Application
1) cDNA synthesis reaction;
2) Polysome isolation;
3) In vitro translation;
4) In vitro transcription of SP6 or T7 RNA polymerase
5) In vitro transcription with cell-free extract.
Use attention
1) The pH range of inhibitory activity is wide, and the maximum activity is shown at pH 7~8;
2) Foaming or strong stirring (Vortex, etc.) will cause inactivation;
3) Does not inhibit RNase H activity.
Use case
Synthesis of the first strand of cDNA.
1) Prepare the following reaction solution in a microcentrifuge tube, the total volume is 20μl
Reagent | Volume | Final concentration |
Total RNA or Poly(A)RNA | 1-5μg | — |
5×MMLV RT Buffer | 4μl | 1× |
dNTP Mixture(25mM ) | 0.4μl | 0.5mM |
Oligo dT or Random Primer (20μM) | 1μl | 1μM |
RNasin(40U/μl) | 1μl | 2U/μl |
MMLV Reverse Transcriptase(200U/μl) | 1μl | — |
Ultra-pure water | Up to 20μl | — |
Total capacity | 20μl | — |
2) Stir gently.
3) After placing it at room temperature for 10 minutes, move it into a 42°C temperature bath.
4) Incubate at 42°C for 1 hour.
5) Move the sample into ice and cool for 2 minutes.
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| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
| 分析证书 | R292575 |